• Korean Journal of Agricultural Science
• /
• v.27 no.1
• /
• pp.33-38
• /
• 2000

This study was carried out to investigate the characterizations of $\kappa$ -CN genes of milk protein. In order to find out the characterizations of $\kappa$ -CN genes, the nucleotide sequences of 5'-flanking regions of $\kappa$ -CN genes were analyzed by PCR(polymerase chain reaction) technique with specific primers in order to investigate the characterizations of these promoters in Korean Native cattle and Holstein. PCR products obtained 349bp fragment and the nucleotide sequences of Korean Native cattle and Holstein of $\kappa$ -CN gene S'-flanking region was analyzed to -82bp from -431bp. On the comparison of each breed, Holstein substituted T$\rightarrow$C at -386bp, and -241bp(T) and -192bp(C) existed, but Korean Native cattle was deleted. Also, Korean Native cattle was existed T at -183bp but Holstein was not. The homology analysis of between Korean Native cattle and Holstein was showed 98.9% homology for $\kappa$ -CN promoter.

• Korean Journal of Plant Tissue Culture
• /
• v.21 no.6
• /
• pp.353-356
• /
• 1994

Chimeric kappa chain and gamma chain cDNA clones (pCKS2 and pCHS2) of a monoclonal antibody specific for pre-S2 surface antigen of human hepatitis-B virus were ligated into Xbal site of plant expression vector pBKS-1. Plasmid DNA containing each of the chimeric gene were then mobilized from E, coli to Agrobacterium tumefaciens strain LBA4404. The chimeric antibody genes were then introduced into tobacco by Ti plasmid-mediated transformation. The putative Transformants were selected on medium containing kamaycin sulfate. Shoots that formed on shoot induction medium were analyzed by Western blot analysis for the expression of kappa-chain or gamma-chain genes. The Western blot analyses clearly showed that the introduced genes were stably expressed in transgenic plants.

• Molecules and Cells
• /
• v.41 no.12
• /
• pp.1008-1015
• /
• 2018

$I{\kappa}B$, a cytoplasmic inhibitor of nuclear factor-${\kappa}B$ ($NF-{\kappa}B$), is reportedly degraded via the proteasome. However, we recently found that long-term incubation with proteasome inhibitors (PIs) such as PS-341 or MG132 induces $I{\kappa}B{\alpha}$ degradation via an alternative pathway, lysosome, which results in $NF-{\kappa}B$ activation and confers resistance to PI-induced lung cancer cell death. To enhance the anti-cancer efficacy of PIs, elucidation of the regulatory mechanism of PI-induced $I{\kappa}B{\alpha}$ degradation is necessary. Here, we demonstrated that PI up-regulates nuclear factor (erythroid-derived 2)-like 2 (Nrf2) via both de novo protein synthesis and Kelch-like ECH-associated protein 1 (KEAP1) degradation, which is responsible for $I{\kappa}B{\alpha}$ degradation via macroautophagy activation. PIs increased the protein level of light chain 3B (LC3B, macroautophagy marker), but not lysosome-associated membrane protein 2a (Lamp2a, the receptor for chaperone-mediated autophagy) in NCI-H157 and A549 lung cancer cells. Pretreatment with macroautophagy inhibitor or knock-down of LC3B blocked PI-induced $I{\kappa}B{\alpha}$ degradation. PIs up-regulated Nrf2 by increasing its transcription and mediating degradation of KEAP1 (cytoplasmic inhibitor of Nrf2). Overexpression of dominant-negative Nrf2, which lacks an N-terminal transactivating domain, or knock-down of Nrf2 suppressed PI-induced LC3B protein expression and subsequent $I{\kappa}B{\alpha}$ degradation. Thus, blocking of the Nrf2 pathway enhanced PI-induced cell death. These findings suggest that Nrf2-driven induction of LC3B plays an essential role in PI-induced activation of the $I{\kappa}B$/$NF-{\kappa}B$ pathway, which attenuates the anti-tumor efficacy of PIs.

• Bulletin of the Korean Chemical Society
• /
• v.32 no.5
• /
• pp.1692-1696
• /
• 2011

The reactions of 1H-1,2,4-triazole (Htr) with $MX_2$ ($ZnCl_2$ for 1; $CdBr_2$ for 2) resulted in two coordination polymers, [Zn(tr)Cl]$_n$ (1) and $[Cd(Htr)_2Br_2]_n$ (2). The structural analyses indicate that 1 and 2 feature a 2D layer and 1D triple chain, respectively. In 1, neighouring Zn atoms are connected by ${\mu}_3-1$ ${\kappa}N$: 2 ${\kappa}N$: $4{\kappa}N-tr^-$ anionic ligand into 6- and 16-membered rings, further grow into a 2D sheet. Cd atoms in 2 are bonded by two ${\mu}_2-Br^-$ bridges and neutral ${\mu}_2$-1 ${\kappa}N$: 2 ${\kappa}N$-Htr to form a 1D triple chain. The fluorescent characterizations of 1, 2 and the free Htr ligand feature simlilar emission peakes at 444, 446 and 423 nm respectively, which can be assigned to intra-ligand ${\pi}-{\pi}^*$ transition of (H)tr. The energy gaps of 5.90 eV for 1, 5.16 eV for 2, and 5.93 eV for Htr suggest that the compounds behave as insulators.

• YAKHAK HOEJI
• /
• v.57 no.6
• /
• pp.426-431
• /
• 2013

In our previous studies, 6-hydroxy-7-methoxychroman-2-carboxylic acid N-phenylamide (KL-1156) was identified as a good inhibitor of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activation. In continuation of our study, we describe the structure-activity relationship of chroman derivatives containing N-arylalkyl groups and their NF-${\kappa}B$ inhibitory activities. In addition, inhibitory effects of cell proliferation are evaluated against human cancer cell lines (NCI-H23 and PC-3). The most active compounds 3i and 3j contained diphenylethyl and diphenylpropyl side chain on amide nitrogen.

• Journal of Periodontal and Implant Science
• /
• v.41 no.3
• /
• pp.157-163
• /
• 2011

Purpose: Curcumin is known to exert numerous biological effects including anti-inflammatory activity. In this study, we investigated the effects of curcumin on the production of interleukin-6 (IL-6) by murine macrophage-like RAW 264.7 cells stimulated with lipopolysaccharide (LPS) from Prevotella intermedia, a major cause of inflammatory periodontal disease, and sought to determine the underlying mechanisms of action. Methods: LPS was prepared from lyophilized P. intermedia ATCC 25611 cells by the standard hot phenol-water method. Culture supernatants were collected and assayed for IL-6. We used real-time polymerase chain reaction to detect IL-6 mRNA expression. $I{\kappa}B-{\alpha}$ degradation, nuclear translocation of NF-${\kappa}B$ subunits, and STAT1 phosphorylation were characterized via immunoblotting. DNA-binding of NF-${\kappa}B$ was also analyzed. Results: Curcumin strongly suppressed the production of IL-6 at both gene transcription and translation levels in P. intermedia LPS-activated RAW 264.7 cells. Curcumin did not inhibit the degradation of $I{\kappa}B-{\alpha}$ induced by P. intermedia LPS. Curcumin blocked NF-${\kappa}B$ signaling through the inhibition of nuclear translocation of NF-${\kappa}B$ p50 subunit. Curcumin also attenuated DNA binding activity of p50 and p65 subunits and suppressed STAT1 phosphorylation. Conclusions: Although further study is required to explore the detailed mechanism of action, curcumin may contribute to blockade of the host-destructive processes mediated by IL-6 and appears to have potential therapeutic values in the treatment of inflammatory periodontal disease.

• BMB Reports
• /
• v.48 no.10
• /
• pp.559-564
• /
• 2015

We investigated the effects of mangiferin on the expression and activity of metalloproteinase (MMP)-9 and the invasion of tumor necrosis factor (TNF)-$\alpha$-stimulated human LNCaP prostate carcinoma cells. Reverse-transcription polymerase chain reaction (RT-PCR) and western blot analysis showed that mangiferin significantly reversed TNF-$\alpha$-induced mRNA and protein expression of MMP-9 expression. Zymography data confirmed that stimulation of cells with TNF-$\alpha$ significantly increased MMP-9 activity. However, mangiferin substantially reduced the TNF-$\alpha$-induced activity of MMP-9. Additionally, a matrigel invasion assay showed that mangiferin significantly reduced TNF-$\alpha$-induced invasion of LNCaP cells. Compared to untreated controls, TNF-$\alpha$-stimulated LNCaP cells showed a significant increase in nuclear factor-${\kappa}B$ (NF-${\kappa}B$) luciferase activity. However, mangiferin treatment markedly decreased TNF-$\alpha$-induced NF-${\kappa}B$ luciferase activity. Furthermore, mangiferin suppressed nuclear translocation of the NF-${\kappa}B$ subunits p65 and p50. Collectively, our results indicate that mangiferin is a potential anti-invasive agent that acts by suppressing NF-${\kappa}B$-mediated MMP-9 expression.

• Journal of Korean Neurosurgical Society
• /
• v.56 no.5
• /
• pp.375-382
• /
• 2014

Objective : To assess the effect of bone marrow mononuclear cells (BMMNCs) transplantation in the expression of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) in spinal cord injury (SCI) in rats. Methods : BMMNCs were isolated from tibia and femur by a density gradient centrifugation. After establishment of acute transection SCI, rats were divided into experiment (BMMNCs), experiment control (0.1 M PBS infused) and sham surgery groups (laminectomy without any SCI). Locomotor function was assessed weekly for 5 weeks post-injury using BBB locomotor score and urinary bladder function daily for 4 weeks post-injury. Activity of NF-${\kappa}B$ in spinal cord was assessed by immunohistochemistry and reverse transcriptase polymerase chain reaction. Results : At each time point post-injury, sham surgery group had significantly higher Basso, Beattie, Bresnahan locomotor and urinary bladder function scores than experiment and experiment control group (p<0.05). At subsequent time interval there were gradual improvement in both experiment and experiment control group, but experiment group had higher score in comparison to experiment control group (p<0.05). Comparisons were also made for expression of activated NF-${\kappa}B$ positive cells and level of NF-${\kappa}B$ messenger RNA in spinal cord at various time points between the groups. Activated NF-${\kappa}B$ immunoreactivity and level of NF-${\kappa}B$ mRNA expression were significantly higher in control group in comparison to experiment and sham surgery group (p<0.05). Conclusion : BMMNCs transplantation attenuates the expression of NF-${\kappa}B$ in injured spinal cord tissue and thus helps in recovery of neurological function in rat models with SCI.

• Biomolecules & Therapeutics
• /
• v.26 no.6
• /
• pp.560-567
• /
• 2018

In the present study, we tried to examine whether resveratrol regulates the expression of matrix metalloproteinases (MMPs) through affecting nuclear factor-kappa B ($NF-{\kappa}B$) in articular chondrocytes. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-${\beta}$ ($IL-1{\beta}$)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. Effect of resveratrol on $IL-1{\beta}$-induced secretion of MMP-3 was investigated in rabbit articular chondrocytes using western blot analysis. To elucidate the action mechanism of resveratrol, effect of resveratrol on $IL-1{\beta}$-induced $NF-{\kappa}B$ signaling pathway was investigated in SW1353, a human chondrosarcoma cell line, by western blot analysis. The results were as follows: (1) resveratrol inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) resveratrol reduced the secretion of MMP-3; (3) resveratrol inhibited $IL-1{\beta}$induced activation (phosphorylation) of inhibitory kappa B kinase (IKK), and thus phosphorylation and degradation of inhibitory kappa $B{\alpha}$ ($I{\kappa}B{\alpha}$); (4) resveratrol inhibited $IL-1{\beta}$-induced phosphorylation and nuclear translocation of $NF-{\kappa}B$ p65. This, in turn, led to the down-regulation of gene expression of MMPs in SW1353 cells. These results suggest that resveratrol can regulate the expression of MMPs through affecting $NF-{\kappa}B$ by directly acting on articular chondrocytes.

• The Korean Journal of Physiology and Pharmacology
• /
• v.21 no.4
• /
• pp.449-456
• /
• 2017

Beauvericin (BEA), a cyclic hexadepsipeptide produced by the fungus Beauveria bassiana, is known to have anti-cancer, anti-inflammatory, and anti-microbial actions. However, how BEA suppresses macrophage-induced inflammatory responses has not been fully elucidated. In this study, we explored the anti-inflammatory properties of BEA and the underlying molecular mechanisms using lipopolysaccharide (LPS)-treated macrophage-like RAW264.7 cells. Levels of nitric oxide (NO), mRNA levels of transcription factors and the inflammatory genes inducible NO synthase (iNOS) and interleukin (IL)-1, and protein levels of activated intracellular signaling molecules were determined by Griess assay, semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR), luciferase reporter gene assay, and immunoblotting analysis. BEA dose-dependently blocked the production of NO in LPS-treated RAW264.7 cells without inducing cell cytotoxicity. BEA also prevented LPS-triggered morphological changes. This compound significantly inhibited nuclear translocation of the $NF-{\kappa}B$ subunits p65 and p50. Luciferase reporter gene assays demonstrated that BEA suppresses MyD88-dependent NF-${\kappa}B$ activation. By analyzing upstream signaling events for $NF-{\kappa}B$ activation and overexpressing Src and Syk, these two enzymes were revealed to be targets of BEA. Together, these results suggest that BEA suppresses $NF-{\kappa}B$-dependent inflammatory responses by suppressing both Src and Syk.